Modular Protein Cages for Size-Selective RNA Packaging in Vivo

Abstract

Protein cages have recently emerged as an important platform for nanotechnology development. Of the naturally existing protein cages, viruses are among the most efficient nanomachines, overcoming various barriers to achieve component replication and efficient self-assembly in complex biological milieu. We have designed an artificial system that can carry out the most basic steps of viral particle assembly in vivo. Our strategy is based on patchwork capsids formed from Aquifex aeolicus lumazine synthase and a circularly permuted variant with appended cationic peptides. These two-component protein containers self-assemble in vivo, capturing endogenous RNA molecules in a size-selective manner. By varying the number and design of the RNA-binding peptides displayed on the lumenal surface, the length of guest RNA can be further controlled. Using a fluorescent aptamer, we also show that short-lived RNA species are captured by the protein cage. This platform has potential as a model system for investigating virus assembly, as well as developing RNA regulation or sampling tools to augment biotechnology.

A Multifunctional Peptide‐Conjugated AIEgen for Efficient and Sequential Targeted Gene Delivery into the Nucleus

“…Moreover, it has been proved that a multidomain designer vector with complex and multifunctional peptides might retain the individual functionality of its constituents through rational arrangement . Hilvert and co‐workers developed a simple and efficient modular system for the in vivo encapsulation of RNA with patchwork capsids, RNA‐binding peptides, a nuclear localization signal (NLS), and cationic peptides . However, for efficient and accurate treatment, subcellular organelle targeting, modular design, and real‐time and long‐term tracking should be considered together for oligonucleotide‐based therapies.…”

Structural Insight into Binary Protein Metal–Organic Frameworks with Ferritin Nanocages as Linkers and Nickel Clusters as Nodes

“…Emulating protein function with synthetic molecules remains a great challenge, and therefore constructing materials with proteins as building blocks has been the best way to make use of the advantageous properties of proteins. Ubiquitous protein nanocages have received considerable attention due to their valuable properties . Recently, Tezcan et al.…”

“…Ubiquitous protein nanocages have received considerable attentiond ue to their valuablep roperties. [20][21][22][23][24][25] Recently,T ezcan et al utilized pseudo spherical ferritin molecules with eight zinc ions decorated on their outer surfacea sh uge nodes and synthetic ditopic hydroxamica cida sl inkers to construct protein MOFs where ferritin moleculesa rrangei nto the desired body-centered cubic (bcc) lattice. [26,27] By taking advantage of the metalc oordination, the Te zcan group also rebuilts ingle ferritin molecules with cupper and controlled the expansion and contraction of polymer-bonded ferritin crystalsw ith different metals.…”

A Multifunctional Peptide‐Conjugated AIEgen for Efficient and Sequential Targeted Gene Delivery into the Nucleus

“…[8] Moreover,ithas been proved that amultidomain designer vector with complex and multifunctional peptides might retain the individual functionality of its constituents through rational arrangement. [10] However,f or efficient and accurate treatment, subcellular organelle targeting, modular design, and real-time and long-term tracking should be considered together for oligonucleotide-based therapies.Amultifunctional peptide-conjugated vector for efficient gene delivery as well as real-time and long-term tracking in the targeted nucleus is highly desirable. [10] However,f or efficient and accurate treatment, subcellular organelle targeting, modular design, and real-time and long-term tracking should be considered together for oligonucleotide-based therapies.Amultifunctional peptide-conjugated vector for efficient gene delivery as well as real-time and long-term tracking in the targeted nucleus is highly desirable.…”

“…[9] Hilvert and co-workers developed as imple and efficient modular system for the in vivo encapsulation of RNAw ith patchwork capsids,R NA-binding peptides,anuclear localization signal (NLS), and cationic peptides. [10] However,f or efficient and accurate treatment, subcellular organelle targeting, modular design, and real-time and long-term tracking should be considered together for oligonucleotide-based therapies.Amultifunctional peptide-conjugated vector for efficient gene delivery as well as real-time and long-term tracking in the targeted nucleus is highly desirable.…”